铜绿假单胞菌的耐药性及其耐氟喹诺酮机制的研究

目的：了解铜绿假单胞菌的耐药性及其耐氟喹诺酮的机制。方法：用纸片扩散法测定626株铜绿假单胞菌对11种抗生素的耐药性。琼脂稀释法测定40株耐环丙沙星的铜绿假单胞菌对环丙沙星和左氧氟沙星的最低抑菌浓度（MIC），聚合酶链反应－限制性长度多态性分析检测耐环沙星的铜绿假单胞菌的gyrA基因和parC基因突变，结果：626株铜绿假单胞菌中耐环丙沙星的180株（28．8％），耐左氧氟沙星的219株（35．0％），40株耐环丙沙星的铜绿假单胞菌有30株（75％）发生gyrA基因83位点突变，26株（65％）发生parC基因87位点突变，两种突变同时发生的25株（62．5％），结论：铜绿假单胞菌的耐药性日趋严重，临床分离的铜绿假单胞菌耐氟喹诺酮的机制大多为药物作用靶位gyrA和parC的基因突变。     

2045份痰培养病原菌分析

本文报告医大附属一院检验科１９９４年￣１９９５年痰培养２０４５例，检出病原菌５７３株，阳性检出率为２８％，前五位细菌是：金黄色葡萄球菌１４８株（２５．８％）、白色念珠菌７６株（１３．２％），铜绿假单胞菌６４株（１１．１％），其他假单胞菌５４株（９．４％），肺炎克雷白氏菌４９株（８．５％），金黄色葡萄球菌对万古霉素、丁胺卡那霉素，环丙沙星、头孢哌酮敏感率在７５％以上，对青霉素Ｇ、红霉素多数耐药、铜绿     

假单胞菌属医院感染176例分析

目的 了解单胞菌医院感染现况,探讨防治对策。方法 对全院６年间医院感染调查病例中临床标本分离出假单胞菌者１７６例进行回顾性分析。结果 共分离出单胞菌１９４株,其中铜绿假单胞菌９６株（４９．４８％）,非铜绿假单胞菌９８株（５０．５２％）,非铜绿假单胞菌中嗜麦芽假单胞菌４６株（２３．７１％）,占第１位,病原菌分布在脓液／分泌液中最多４８．４５％,其次为痰３０．９３％、尿１２．８９％。结论 假单胞菌感染易发生于各种原因导致的免疫机能下降者,在临床标本中分布广泛,且对抗生素的耐药性逐渐增加,应重视假单胞菌在医院感染中的作用。     

耐碳青霉烯类铜绿假单胞菌耐药基因的研究

目的探讨瑞安市人民医院临床分离的耐碳青霉烯类铜绿假单胞菌的耐药特征与基因型分布,为临床合理使用抗生素提供参考依据。方法收集本院2012年1月-2017年12月经VITEK 2全自动微生物分析仪鉴定的铜绿假单胞菌10 288株;分离4 072株耐碳青霉烯类铜绿假单胞菌并进行耐药特征分析;筛选耐碳青霉烯菌210株,采用PCR法进一步分析其基因型:检测外膜通透蛋白Opr-D2基因及5种主要金属β-内酰胺酶基因(IMP-1、IMP-2、VIM-1、VIM-2、SPM)。结果 10 288株铜绿假单胞菌以痰液标本为主,主要分布于重症监护室;4 072株耐碳青霉烯类铜绿假单胞菌对替卡西林/克拉维酸、美罗培南的耐药率相对较高;210株耐碳青霉烯类铜绿假单胞菌中携带有SPM基因的铜绿假单胞菌18株,占8.6%,携带Opr-D2基因54株,外膜蛋白缺失率为74.3%,其余基因型均阴性。结论本院耐碳青霉烯类铜绿假单胞菌的耐药机制主要为外膜蛋白Opr-D2缺失和产SPM型金属酶,临床应结合药敏谱,合理使用抗生素。     

呼吸病区铜绿假单胞菌医院感染的流行病学研究

为了解铜绿假单胞菌医院感染的流行病学。方法 用血清型和抗菌谱联合分型方法，对患者分离株和环境分离株从检出时间，地点或部位进行动态分析。结果 ２１２３份样本共检出铜绿假单胞菌１７６株，分离率为０．８％＿５７．１％，患者之间，患者与环境及医护人员检出的铜绿假单胞菌未见有关联。     

耐碳青酶烯类铜绿假单胞菌相关耐药基因及I型整合酶基因研究

目的研究耐碳青酶烯类铜绿假单胞菌的相关耐药基因及Ⅰ型整合酶基因。方法采用聚合酶链反应（PCR）法对铜绿假单胞菌碳青酶烯类耐药相关的金属β-内酰胺酶IMP、VIM、SPM、GIM基因和外膜蛋白oprD2基因及Ⅰ型整合酶基因等6种主要耐药基因进行检测与分析。结果51株耐亚胺培南和美罗培南铜绿假单胞菌SPM、GIM金属酶基因检测均阴性,16株IMP和5株VIM型金属酶基因阳性,14株耐亚胺培南铜绿假单胞菌oprD2基因阳性,其余37株铜绿假单胞菌oprD2基因阴性,7株产金属酶并同时伴有膜孔蛋白oprD2基因缺失,49株Ⅰ型整合酶基因intⅠ1阳性。结论膜孔蛋白oprD2基因缺失是铜绿假单胞菌对碳青酶烯类抗菌药物耐药的主要原因,其次是产生金属酶,Ⅰ型整合酶基因广泛存在于铜绿假单胞菌中,提示要加强对耐药基因在病原菌种属间传播和扩散的监测工作。     

ＩＣＵ 多重耐药铜绿假单胞菌感染肺炎的治疗及 降钙素原对抗菌药物使用的指导意义

摘要：目的　探讨ＩＣＵ 多重耐药铜绿假单胞菌感染肺炎的临床治疗对策，以及降钙素原对临床抗菌药物使 用的指导意义。方法　选取２０１２年１月至２０１７年１２月温州医科大学附属第二医院ＩＣＵ 病房收治的经痰液 样本分离出的铜绿假单胞菌感染肺炎患者１４２例，根据药敏试验结果分为非多重耐药菌种和多重耐药菌， 每组７１例，对两组患者分别使用抗菌药物进行治疗，对其临床效果进行探讨。同时，在治疗前及治疗后 ７ｄ、１４ｄ、２１ｄ对７１例ＩＣＵ 多重耐药铜绿假单胞菌感染肺炎患者降钙素原进行检测，对比使用一种抗菌 药物和两种及以上抗菌药物在治疗前后的降钙素原，以及治疗后炎性指标的情况。结果　非多重耐药菌种 采用一种抗菌药物治疗的效果低于两种及以上抗菌药物治疗效果， 两组相比差异无统计学意义（χ ２ ＝ ０．１３７，犘＞０．０５），多重耐药菌种采用一种抗菌药物治疗的效果低于两种及以上抗菌药物治疗效果，两组 相比差异有统计学意义（χ ２＝２１．３８４，犘＜０．０５）；观察组与治疗组患者治疗前ＰＣＴ 检测结果差异无统计 学意义（狋＝０．２５９，犘＞０．０５）；治疗后，两组ＰＣＴ 检测结果均有改善，观察组改善优于对照组，两组相比 差异有统计学意义（狋＝４．５７２、６．８３９、１１．２８７，犘＜０．０５）；治疗结束后，观察组ＣＲＰ、白细胞计数、ＩＬ ６ 均低于对照组，两组差异有统计学意义（狋＝１９．６５２、７．３８４、２８．３９１，犘＜０．０５）。结论　ＩＣＵ 铜绿假单胞 菌感染肺炎患者应选用两种及以上抗菌药物进行治疗，ＰＣＴ 的检测对ＩＣＵ 多重耐药铜绿假单胞菌感染肺炎 抗菌药物的使用起到指导性的作用。 关键词：ＩＣＵ；铜绿假单胞菌；降钙素原；抗菌药物；多重耐药；肺炎；治疗 中图分类号：Ｒ５６３．１、Ｒ９７８．１、Ｒ４６６．１１　　文献标识码：Ａ　　文章编号：１００９ ６６３９ （２０１９）０３ ０１７８ ０４     

耐碳青霉烯类铜绿假单胞菌产金属β-内酰胺酶的检测及耐药性分析

目的了解医院耐碳青霉烯类铜绿假单胞菌产金属β-内酰胺酶(MBL)情况及其对临床常用抗菌药物的耐药性,为临床抗感染治疗提供依据。方法采用E-test检测铜绿假单胞菌是否产生MBL,用BD Phoenix 100全自动微生物测定铜绿假单胞菌对常用15种抗菌药物的耐药性。结果在分离的189株铜绿假单胞菌中耐碳青霉烯类的有54株(28.6%),其中产MBL的有13株(24.1%);耐碳青霉烯类铜绿假单胞菌与碳青霉烯类敏感铜绿假单胞菌对大多数常用抗菌药物的敏感性差异有统计学意义。结论医院耐碳青霉烯类的铜绿假单胞菌检出率较高,多药耐药现象较严重,产MBL是铜绿假单胞菌对碳青霉烯类药物耐药的主要原因之一,临床医师应参考微生物实验室的药敏结果,合理选用抗菌药物。     

呼吸与危重症医学病房分离的碳青霉烯耐药铜绿假单胞菌的分子流行病学

目的了解呼吸与危重症医学病房分离的碳青霉烯耐药铜绿假单胞菌的分子流行病学特点。方法收集陆军军医大学大坪医院2018年1-12月呼吸与危重症医学病房分离的20株碳青霉烯耐药铜绿假单胞菌,检测其对常用抗菌药物的敏感性,通过聚合酶链反应(Polymerase chain reaction,PCR)扩增主要的碳青霉烯酶基因(bla_(KPC-2)、bla_(IMP)、bla_(VIM)、bla_(SPM)、bla_(NDM-1)和bla_(OXA-48)),利用脉冲场凝胶电泳(Pulsed field gel electrophoresis,PFGE)对20株碳青霉烯耐药铜绿假单胞菌进行同源性分析。结果呼吸与危重症医学病房分离的20株碳青霉烯耐药铜绿假单胞菌对10种常用抗菌药物的耐药率均40.0%; 20株碳青霉烯耐药铜绿假单胞菌中有6株携带bla_(VIM)基因,6株携带bla_(SPM)基因,2株携带bla_(KPC)基因,剩余6株未携带耐药基因; PFGE结果显示,20株碳青霉烯耐药铜绿假单胞菌可分为A～F共6个谱型,其中E型占35%,C型占20%。结论医院呼吸与危重症医学病房分离的碳青霉烯耐药铜绿假单胞菌主要以产SPM型和VIM型碳青霉烯酶为主,存在部分耐碳青霉烯铜绿假单胞菌的克隆传播,临床上应严格做好耐药菌的防控工作,避免耐碳青霉烯铜绿假单胞菌在呼吸与危重症医学病房感染暴发流行。     

铜绿假单胞菌的耐药性分析

目的：了解本院2010年1-12月铜绿假单胞菌的分离情况及耐药特点。方法：从2010年1-12月老年住院患者的痰液、分泌物等标本中分离出102株铜绿假单胞菌,统计其样本来源、科室分布、对不同抗菌药物的敏感性。结果：在检出的102株铜绿假单胞菌中58株来自呼吸内科,占56.86%;其次为神经外科,占13.73%,比较敏感的抗菌药物依次是多粘菌素（100%）、亚胺培南（82.36%）、哌拉西林/他唑巴坦（74.51%）。结论：铜绿假单胞菌为非发酵菌中的假单胞菌属,很容易造成机会性感染,临床应合理选用抗菌药物,从而减少耐药菌株的产生,有效抑制铜绿假单胞菌耐药率过快增长,同时应不断检测抗菌药物的敏感性,为临床合理用药提供科学依据。     

Excretion products of algae and their occurrence in Solar Lake. Taba,Egypt

The present study aimed to investigate the nature of organic compounds liable to be released by blue-green algae (cyanobacteria) and organic compounds detected in the Solar Lake in Taba, Egypt. The liquid-liquid extraction method and liquid chromatography technique were applied for extraction, clean-up and separation of the organics. Gas chromatography/mass spectrometry (GC/MS) was used to carry out qualitative analysis. The most dominant species in the algal samples collected from Solar lake in Taba are Aphanotheca stagnina, Synechococcus aeruginosus, Oscillatoria limnetica and Microcoleus chthonoplastes. n-Alkane hydrocarbons that ranged from C12 to C25 were identified in lake water samples, most of them were derived from algal species. Several fatty acids were isolated from water samples with predominant tridecanoic acid (C13), tetradecanoic acid (C14) and hexadecanoic acid (C16). Meanwhile, the identification of fatty acids from algal suspension samples indicated the predominance of hexadecanoic acid (C16) and heptadecanoic acid (C17), with carbon number that ranged from C10 to C17. The dominant compounds were C15, C17, C17:1, C19 n-alkanes. C13, C15, C16 fatty acids, alkyl esters of propyl decanoate, esters of benzenedicarboxlate, diphenyl amine, geosmin and 2-methyl isoborneol (2-MIB) in water and in algal suspension samples were probably enough to distinguish the cyanobacterial mat community of a saline lake shore.     

食品中沙门菌细胞脂肪酸组分的气相色谱分析

目的：建立沙门菌属全细胞脂肪酸气相色谱指纹图谱，为沙门菌的鉴定提供依据。方法：对从食品中分离的6种沙门菌在相同的条件下培养，离心收集细胞，提取脂肪酸并甲基化，通过TLC来分离纯化甲基酯，通过气相色谱测定脂肪酸的组成。结果：沙门菌属中的细菌在脂肪酸的组成上主要含C12、C16、C18、C19。不同种的脂肪酸组分表现出一定的差异，地方菌株与模式菌株具有相同的脂肪酸组分。结论：该方法灵敏度高、重现性好，准确性强可以对食品中的沙门菌进行快速鉴定。     

两种铜绿假单胞菌同源性分析方法比较

目的了解脂肪酸气相色谱分析方法能否应用于铜绿假单胞菌的同源性分析。方法收集某院2008年1—12月重症监护室住院患者痰标本分离的26株多重耐药铜绿假单胞菌,分别应用脉冲场凝胶电泳(PFGE)和脂肪酸气相色谱分析方法进行同源性分析,比较两组结果。结果 PFGE结果表明,26株菌有6个克隆;脂肪酸气相色谱分析显示,26株菌存在4个克隆,符合率仅26.92%(7/26)。结论脂肪酸气相色谱分析方法在菌株同源性判断的准确性上与PFGE分型有差距,其应用于铜绿假单胞菌的同源性分析仍需谨慎。     

唐菖蒲伯克霍尔德菌菌体脂肪酸成分测定与分析

目的对唐菖蒲伯克霍尔德菌不同致病型菌株的菌体脂肪酸成分进行测定和分析。方法利用气相色谱法测定唐菖蒲伯克霍尔德菌不同致病型菌株的菌体脂肪酸成分,并利用MIDI-FAME方法进行分析。结果研究发现椰毒假单胞菌的菌株与唐菖蒲伯克霍尔德菌中其他致病型菌株的脂肪酸成分非常相似,从而进一步确定了它的分类地位。另外还发现C16∶0.2OH、C18∶1.2OH和C16∶1.2OH与椰毒假单胞菌酵米面亚种的米酵菌酸的产生具有一定的联系。结论由脂肪酸成分分析确认了椰毒假单胞菌的分类地位,为进一步研究它的产毒机制和致病机理积累资料。     

Epidemiology of Pseudomonas aeruginosa infection and the role of contamination of the environment in a cystic fibrosis clinic

In order to identify the possible reservoirs and routes of cross-infection with Pseudomonas aeruginosa, samples from patients, staff, and the environment of a cystic fibrosis centre and two control wards at an infectious disease clinic were collected during a two-week period in 1980. All the Ps. aeruginosa strains were phage and serotyped. Ps. aeruginosa was isolated from 90 (51%) of the cystic fibrosis patients and most belonged to the 0-3/9 complex, characteristic of strains from patients in the centre. Some of the patients were able to spread Ps. aeruginosa into the air and to their hands by coughing, and Ps. aeruginosa in dried sputum could survive for at least one week. Strains of the same epidemiological types as found in the cystic fibrosis patients were isolated from sinks, soap, baths, toys, tables, brushes, cloths, and air in the clinic. In contrast, Ps. aeruginosa of the same epidemiological types were only found in a few of the sinks in one of the control wards where a few cystic fibrosis patients were regularly treated in isolation cubicles. The precautions employed to prevent future cross-infection include segregation of Ps. aeruginosa-infected from non-infected patients in separate wards and arranging for visits on separate days in the out-patients clinic. The survival of cystic fibrosis patients treated in the centre is much longer than those treated outside the centre despite the problems of cross-infection.     

Low level of bacterial contamination of mist tents used in home treatment of cystic fibrosis patients

Mist tents are recommended by the Stockholm cystic fibrosis (CF) centre for small children with CF. Daily disinfection of some parts of the tent with 2% acetic acid is recommended, and for other parts boiling water followed by air-drying without rinsing. The plastic tent is discarded each day. We have studied whether these prescribed routines are followed by the patients and whether they are sufficient to prevent bacterial contamination. The mist tent equipment of 20 CF patients (mean age 7 years, range 1-15 years), two of whom were chronically colonized with Pseudomonas aeruginosa, were investigated. All patients were visited at home in the morning after 6-12 hours aerosol therapy. Liquid from the nebulizing chambers and swabs from the aerosol tube were examined by culture on four different media. Seventeen of 20 patients claimed that they cleaned and disinfected the tubes every day, two patients every other day and one once a week. Seventeen of 19 claimed they cleaned and disinfected the chambers daily, one once a week and, one twice a week. No or insignificant growth was found in 16/20 aerosol tubes: moulds in three, Pseudomonas species in one. Twelve of 19 chambers showed no or insignificant growth: moulds or yeasts were present in three and Pseudomonas sp. in four. In four of the seven patients moulds or yeasts and/or Pseudomonas sp. grew both from chambers and from aerosol tubes; in the remaining three only from chambers. None of these seven patients had followed our prescribed cleaning and disinfection recommendations, the other 13 claimed they had. Of the patients whose equipment yielded Pseudomonas sp, none was colonized with these strains, although one had P. aeruginosa. We conclude that our disinfection recommendations are adequate when followed. However, our disinfection recommendations concerning the nebulizing chamber had not been followed satisfactorily. The different forms of non-compliance would not have been detected without a home visit, emphasizing the importance of such visits. The importance of drying the equipment and of using the correct concentration of acetic acid is stressed.     

脂肪组织和血浆标本中脂肪酸的气相色谱分析

以正十三碳酸(C13:0)和正十七碳酸(C17:0)作内标物,分别对脂肪组织和血浆中总脂肪酸、游离脂肪酸进行气相色谱分析。采用4:1甲醇-苯和氯化乙酰对样品中总脂肪酸同时作抽提和酯化,其酯化率分别是103.5％(内标物C13:0)和102.9％(内标物C17:0);对各次加入量不同的月桂酸、豆蔻酸、硬脂酸和花生酸的回收串为100.0～105.0％,重复性试验中各脂肪酸CV为1.14～8.47％。采用正已烷抽提血浆中游离脂肪酸,并以14％BF_3使甲酯化,其酯化率为103.0％(内标物C17.0),对各次加入量不同的月桂酸、豆蔻酸,硬脂酸和花生酸的回收率是82.0～105.0％。重复性试验中各脂肪酸CV为0.15～5.94％。总脂肪酸法尤为简便、迅速和可靠。     

A new Pseudomonas vaccine: preliminary trial on human volunteers.

Fifteen healthy volunteers were given three weekly subcutaneous injections of a new polyvalent Pseudomonas aeruginosa vaccine (PEV-01). Four doses - 1-0 RHD (manufacturer''s recommended human dose), 0-75 RHD, 0-5 RHD and 0-1 RHD - were used in separate groups of volunteers. Blood samples taken before each of the injections and one taken 7 days after the last injection were examined for immune response to the vaccine and for possible adverse clinical, biochemical and haematological effects. Raised titres of antibody in serum of volunteers given 0-5-1-0 RHD vaccine were shown, often by the seventh day, in passive haemagglutination tests against all of the 16 serotypes of Ps. aeruginosa represented in the vaccine; serum from volunteers who received 0-1 RHD usually showed a reduced antibody titre. Tests of mouse protection by serum against intraperitoneal challenge with Ps. aeruginosa P14 showed increased titres of mouse protective antibody in the blood of volunteers given 1-0, 0-75 or 0-5 RHD of vaccine but a reduced mouse-protective titre in two out of three sera from volunteers given 0-1 RHD vaccine. There was a suggestion of enhanced phagocytic ingestion and intracellular killing of two strains of Ps. aeruginosa by the blood of vaccinated volunteers, and more definite enhancement of ingestion of inert latex particles, which were less well ingested than were the bacterial cells by phagocytes from unvaccinated volunteers. Apart from slight or moderate local reactions and a transient rise of temperature in some volunteers, there were no clinical, biochemical or haematological abnormalities in the vaccinated volunteers.     

Incidence of Pseudomonas aeruginosa in recreational and hydrotherapy pools

Pseudomonas aeruginosa remains an important agent of opportunistic infection in patients, particularly those with respiratory complications and burns. One natural niche of this organism is water and water-associated facilities, hence the aim of this study was to examine specimens from recreational and hydrotherapy pools in Northern Ireland over a two-year period. Water specimens (n = 3,510) were obtained from three amenity categories, namely, 13 hydrotherapy pools (specimen number [n] = 323), 51 Jacuzzis/spas (n = 1,397) and 68 swimming pools (n = 1,790). Specimens (100 ml) were filtered through a cellulose acetate (0.45 micron pore size) gridded filter and the membrane was placed on Pseudomonas CFC agar (Oxoid CM559 + SR103) and incubated at 37 degrees C for 48 +/- 2 h. Colonies that clearly showed pyocyanin production or met other identification criteria were considered P. aeruginosa. Of the amenities examined 4/13 hydrotherapy pools (30.8%), 37/51 Jacuzzis/spas (72.5%) and 26/68 swimming pools (38.2%) were positive for P. aeruginosa. The most heavily contaminated amenity category was the Jacuzzi/spa, where 34.7% and 12% of private and public sites respectively were positive for P. aeruginosa at a level of greater than 1,000 cfu 100 ml-1. Approximately twice as many samples were positive in private Jacuzzis/spas compared to publicly operated facilities. There was a similar trend with respect to public and private hydrotherapy pools, though bacterial counts did not exceed 1,000 cfu 100 ml-1. Recreational and therapeutic amenities involving the use of water may be a potential source of P. aeruginosa for susceptible patient groups, including patients with cystic fibrosis and bronchiectasis. This may vary depending on amenity type and public/private ownership of such amenities.     

呼吸道感染病原菌分布及耐药性调查

目的：了解我院呼吸道感染病原菌谱及耐药性。方法：对我院2006年1月～9月临床送检的痰及咽拭子进行细菌培养,采用美国德灵公司Auto Scan-4微生物分析系统进行细菌鉴定和药物敏感试验;采用ATB Fungus进行真菌药物敏感试验。按NCCLS标准判定敏感菌株。结果：共分离到病原菌119株,革兰阴性杆菌占68.07%,真菌占26.05%,革兰阳性球菌仅占5.88%。革兰阴性杆菌以肺炎克雷伯菌、大肠埃希菌、铜绿假单胞菌为主,分别占病原菌总数的36.13%、7.56%、6.72%;真菌以白色假丝酵母菌为主,占真菌总数的54.84%。药敏结果提示,铜绿假单胞菌、肠杆菌属、沙雷氏菌属等均出现多重高比例耐药,真菌也对唑类药物出现强耐药性。结论：最近呼吸道感染病原菌以革兰阴性杆菌和真菌为主,而且出现多重耐药。